Low-Cost Silicone Imaging Casts for Zebrafish Embryos and Larvae

Masselink, Wouter; Wong, Jin Cheng; Liu, Boyin; Fu, Jianzhong; Currie, Peter D.

doi:10.1089/zeb.2013.0897

Cited by 16 publications

(10 citation statements)

References 10 publications

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“…The microstructures integrated into the divots and channels used here are designed to orient and immobilize the larvae without depending entirely on water tension, thus reducing the risk of drying. Divots made using 3D printed molds have previously been used for orientation of embryos for imaging purposes 13 , although the depth of these divots made them inaccessible for microinjection.…”

Section: Discussionmentioning

confidence: 99%

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Ellett

Irimia

2017

JoVE

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Zebrafish have emerged as a powerful model of various human diseases and a useful tool for an increasing range of experimental studies, spanning fundamental developmental biology through to large-scale genetic and chemical screens. However, many experiments, especially those related to infection and xenograft models, rely on microinjection and imaging of embryos and larvae, which are laborious techniques that require skill and expertise. To improve the precision and throughput of current microinjection techniques, we developed a series of microstructured devices to orient and stabilize zebrafish embryos at 2 days post fertilization (dpf) in ventral, dorsal, or lateral orientation prior to the procedure. To aid in the imaging of embryos, we also designed a simple device with channels that orient 4 zebrafish laterally in parallel against a glass cover slip. Together, the tools that we present here demonstrate the effectiveness of photolithographic approaches to generate useful devices for the optimization of zebrafish techniques.

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Section: Discussionmentioning

confidence: 99%

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Ellett

Irimia

2017

JoVE

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“…All OpenSCAD and corresponding STL design files presented here are freely accessible (see Additional files 1, 5) and can be easily shared electronically. As 3D printers are becoming increasingly widespread, especially at universities and companies, this provides access to the developed tools to a wide range of zebrafish users [9]. Finally, the presented designs can serve as templates for other additive or subtractive manufacturing techniques.…”

Section: Discussionmentioning

confidence: 99%

“…To address this requirement, researchers have developed various protocols and tools that facilitate mounting and positioning of zebrafish embryos and larvae [3-9]. However, most solutions are incompatible with microtiter plates, which are the commonly employed sample holder for automated imaging using commercially available microscopes.…”

Section: Introductionmentioning

confidence: 99%

Generation of orientation tools for automated zebrafish screening assays using desktop 3D printing

Wittbrodt¹,

Liebel

Gehrig

2014

BMC Biotechnol

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BackgroundThe zebrafish has been established as the main vertebrate model system for whole organism screening applications. However, the lack of consistent positioning of zebrafish embryos within wells of microtiter plates remains an obstacle for the comparative analysis of images acquired in automated screening assays. While technical solutions to the orientation problem exist, dissemination is often hindered by the lack of simple and inexpensive ways of distributing and duplicating tools.ResultsHere, we provide a cost effective method for the production of 96-well plate compatible zebrafish orientation tools using a desktop 3D printer. The printed tools enable the positioning and orientation of zebrafish embryos within cavities formed in agarose. Their applicability is demonstrated by acquiring lateral and dorsal views of zebrafish embryos arrayed within microtiter plates using an automated screening microscope. This enables the consistent visualization of morphological phenotypes and reporter gene expression patterns.ConclusionsThe designs are refined versions of previously demonstrated devices with added functionality and strongly reduced production costs. All corresponding 3D models are freely available and digital design can be easily shared electronically. In combination with the increasingly widespread usage of 3D printers, this provides access to the developed tools to a wide range of zebrafish users. Finally, the design files can serve as templates for other additive and subtractive fabrication methods.

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“…This step can be streamlined by automated devices that can pick up and orientate a single larvae for image acquisition and other procedures before dispensing the larvae into an individual well of a multi-well plate [50]. Even smaller-scale manual manipulation and scoring can benefit from using relatively simple microfludic devices or casts to facilitate rapid and reproducible positioning of larvae [51][52][53].…”

Section: Strategiesmentioning

confidence: 99%

Discovery of Novel Therapeutics for Muscular Dystrophies using Zebrafish Phenotypic Screens

Widrick

Kawahara

Alexander

et al. 2019

JND

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The recent availability and development of mutant and transgenic zebrafish strains that model human muscular dystrophies has created new research opportunities for therapeutic development. Not only do these models mimic many pathological aspects of human dystrophies, but their small size, large clutch sizes, rapid ex utero development, body transparency, and genetic tractability enable research approaches that would be inconceivable with mammalian model systems. Here we discuss the use of zebrafish models of muscular dystrophy to rapidly screen hundreds to thousands of bioactive compounds in order to identify novel therapeutic candidates that modulate pathologic phenotypes. We review the justification and rationale behind this unbiased approach, including how zebrafish screens have identified FDA-approved drugs that are candidates for treating Duchenne and limb girdle muscular dystrophies. Not only can these drugs be re-purposed for treating dystrophies in a fraction of the time and cost of new drug development, but their identification has revealed novel, unexpected directions for future therapy development. Phenotype-driven zebrafish drug screens are an important compliment to the more established mammalian, target-based approaches for rapidly developing and validating therapeutics for muscular dystrophies.

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Low-Cost Silicone Imaging Casts for Zebrafish Embryos and Larvae

Cited by 16 publications

References 10 publications

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Generation of orientation tools for automated zebrafish screening assays using desktop 3D printing

Discovery of Novel Therapeutics for Muscular Dystrophies using Zebrafish Phenotypic Screens

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